Vol 5, Issue 1, 2023 (347-356)
http://journal.unpad.ac.id/idjp
*Corresponding author,
e-mail : inesuharyani25@gmail.com (I. Suharyani)
https://doi.org/10.24198/idjp.v5i1.44548
© 2023 I. Suharyani et al
Applcation of Red Dragon Fruit Extract (Hylocereus polyrhizus) as an Antioxidant in
Lip Cream Preparation
Ine Suharyani1,2*, Rindiyani1, Rima Yulia Senja1, Lusi Nurdianti3, Yuniarti Falya1
1Sekolah Tinggi Farmasi Muhammadiyah Cirebon
Cideng Indah, Kertawinangun, Kedawung, Cirebon, Jawa Barat 45133
2Departmen Farmasetika dan Teknologi Farmasi, Fakultas Farmasi, Universitas Padjadjaran,
Jln. Raya Bandung-Sumedang Km. 21 Jatinangor, Kab. Sumedang 45363
3Prodi S1, Universitas Bakti Tunas Husada, Jl. Cilolohan 36 Tasikmalaya, Jawa Barat 46115
Submitted : 15/01/2023, Revised : 22/05/2023,, Accepted : 06/06/ 2023, Published : 08/06/2023
Abstract
Many lip creams contains both synthetic or natural dye. One of the fruit that can
be used as natural dyes in a lip cream preparation is dragon fruit (Hylocereus
polyrhizus) which has an attractive color. In addition, red dragon fruit contains
anthocyanins, a group of flavonoid, that has efficacy as an antioxidants. This
study aim to develope lip cream as cosmetic preparation containing of red dragon
fruit for antioxidant. The extraction was done by Microwave Assisted Extraction
(MAE) using ethanol 95%:aquadest (4:1). Lip cream preparations were made in
3 formulas with varying concentrations of extract, 5% (LCE5), 10% (LCE10) and
15% (LCE 15). The extraction uasing MAE method was yielded the rendemen of
44,12%. Evaluation of the preparations carried out included organoleptic, pH,
homogeneity, and spreadability, while antioxidant activity were carried out using
spectrophotometry UV-Vis with the DPPH (2,2-diphenyl-1-picrylhidrazyl)
method. The results showed that all lip cream formulas fulfill the requirement i.e
organoleptic, pH, homogeneity, and spreadability. The antioxidant activity of
LCE5, LCE10 and LCE15 were 195.26 ± 0.40, 174.94 ± 0,63, and 168.27 ± 0.60,
respectively. All the preparation has medium antioxidant activity but LCE15 has
a stronger antioxidant activity due to the smallest IC50 compared to LCE5 and
LCE10. This result underlying the application of red dragon fruit extract as an
antioxidant agent in lip cream preparation.
Keywords: Antioxidant, Lip cream, Red dragon fruit, IC50
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
348
1. Introduction
Nowadays, there are various forms of
cosmetics on the market, especially cosmetics
for lips, one of which is lip cream. Lip cream is
a cosmetic preparation that has a semi-solid
texture and is used as a lip color (1). In the
process of making cosmetics required
additional materials such as dyes (2). Synthetic
dyes are more widely used in beauty products
because they are cheap, easy to obtain, and
have sharper colors. However, excessive and
continuous use of synthetic dyes can result in
negative health effects such as irritation and
cancer (3).
The efforts to replace the use of synthetic dyes
with natural dyes from plants was elevated, one
of which is the red dragon fruit plant
(Hylocereus polyrhizus). red dragon fruit flesh
(Hylocereus polyrhizus) contains high
concentration of anthocyanin so it can be used
for food, as well as cosmetics both as a coloring
agent and as an antioxidant.
Antioxidants are substances that can withstand
or prevent damage to cells in the body caused
by free radicals. Previous research stated that
the antioxidant activity in red dragon fruit flesh
was higher than in white dragon fruit flesh (4).
To determine the antioxidant activity of a plant,
you can use the DPPH (2,2 diphenyl-1-
picrylhidrazyl) method. The DPPH method is
used because it has a simple procedure to find
out if a compound has antioxidant activity (5).
2. Method
Materials
Red dragon fruit obtained in Cirebon City and
determined in IAIN Syekh Nurjati Cirebon.
Carnauba wax (PT. Pratama Sains Global),
microcrystalline wax (PT. Pratama Sains
Global), setil alkohol (PT. Pratama Sains
Global), castor oil (PT. Pratama Sains Global),
propilen glikol (PT. Pratama Sains Global),
titanium dioksida (PT. Pratama Sains Global),
metil paraben (PT. Pratama Sains Global),
oleum rosae (PT. Pratama Sains Global), tween
80 (PT. Pratama Sains Global), span 80 (PT.
Pratama Sains Global), Vitamin C (PT.
Pratama Sains Global). DPPH from HiMedia
Laboratories, vitamin C from Sigma Aldrich,
while ethanol 96%, and aquadestillata obtained
from Bratachem. All the materials were used
without further purification.
Method
Extraction of red dragon fruit
This study used red dragon fruit (Hylocereus
polyrhizus) with an even red outer skin color
found in Plered, Weru, Cirebon. The red dragon
fruit plant (Hylocereus polyrhizus) was
determined at the Biology Study Program
Laboratory of IAIN Syekh Nur Jati Cirebon to
confirm and confirm the plant samples to be
used.
Red dragon fruit extract was prepared using the
microwave-assisted extraction (MAE) method.
3 kg of red dragon fruit flesh, cut into thin
slices, then put in the oven at 40 oC for 1 day.
After the dragon fruit is dry, it is then soaked in
95% ethanol and distilled water in a ratio of 4:1
for 1 night. The simplicia soaked was put into
the microwave and extracted for 4 minutes,
then filtered to obtain the filtrate using filter
paper and put into the rotary evaporator at 50oC
with a speed of 100 rpm (6). Next, the
concentration of the extract was carried out
using a water bath. The yield obtained is
calculated by the following equation (7):
.......................
.......................................... (1)
Phytochemical screening
Alkaloid
Small amount of sample was diluted with 2 mL
distilled water. Two mililiters solution of 2%
HCl was added, heated for 5 minutes and
filtered. The filtrate is reacted with 3 drops of
Mayer reagent. Positive alkoloid with white
precipitate or yellow precipitate formed (8).
Flavonoid
The sample was diluted with 5 ml of distilled
water, then five drops of magnesium powder
and concentrated HCl were added,
respectively. The formation of red or orange
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
349
color indicates the presence of flavonoids
(Lanisthi dan Febrina, 2015).
Saponin
2 ml of the extract was diluted with 5 ml of
aquadest, then 10 drops of KOH were added
and heated for 5 minutes, shaken for 15
minutes. Positive saponins formed with the
high of 1 ml and stable for 15 minutes (9,10).
Steroid dan Triterpenoid
A piece of extract reacted with 1 ml of
Liberman-Burchard reagent. The appearance of
green indicates steroids, and the red or purple
color indicates the presence of triterpenoids (8).
Tanin
A piece of extract reacted with 2 ml of FeCl3.
The presence of tannins and polyphenolic
compounds indicated with a blackish green or
dark blue (8).
Anthocyanin
A piece of extract was diluted with 2 ml of
water. 2M HCl was added. The prescence of
anthocyanin indicated with the red color (11).
Lip Cream Preparation
Lip cream preparations are made by heating the
fat phase and the water phase in different
containers. The oil phase, namely castor oil,
carnauba wax, microcrystalline wax, and cetyl
alcohol, was heated at 80oC until completely
melted. The water phase, namely distilled
water, propylene glycol, tween 80, and methyl
paraben, was heated at 60 oC. Enter the oil
phase into magnetic stirrer, add the water
phase, and stir until it forms a homogeneous
cream. Add the titanium dioxide and oleum
rosae, then add the red dragon fruit extract, and
continue to stir until homogeneous (12).
Table 1. Formulation of Lip Cream containing Extract (LCE)
For
mula
tion
Extra
ct
(g)
Castor
oil
(mL)
Carnau
ba wax
(g)
Microcr
ystallin
e wax
(g)
Setil
alkoh
ol
(g)
Spa
n
80
(m
L)
Propi
lengl
ikol
(g)
Twee
n 80
(mL)
Metil
parab
en
(g)
Ol.
Rosa
e
(mL)
Distille
d
Water
(mL)
LC
-
12
6
7
5
1,5
10
1,5
0,15
0,05
56,8
LCE
5
5
12
6
7
5
1,5
10
1,5
0,15
0,05
51,8
LCE
10
10
12
6
7
5
1,5
10
1,5
0,15
0,05
46,8
LCE
15
15
12
6
7
5
1,5
10
1,5
0,15
0,05
41,8
Evaluation
Organoleptic observations were carried out
using the five senses to observe the shape, color
and smell of the lip cream preparations (12).
To measure the pH, weigh 1 gram of lip cream
and then dissolve it in a beaker glass with 10
mL of distilled water. Then dip the electrode in
the lip cream until it shows a constant pH value.
Lip pH 4.5–6.5 (13). Homogeneity test, take
enough lip cream then place it between the
object glass and observe whether there are any
granules or not (1).
Spreadability test Take 1 gram of lip cream and
place it in the middle of two flat
*Corresponding author,
e-mail : inesuharyani25@gmail.com (I. Suharyani)
https://doi.org/10.24198/idjp.v5i1.44548
© 2023 I. Suharyani et al
glasses. Add a load weighing 125 g, let it sit
for 1 minute, and note how many diameters
of lip cream are spread. Good spreadability
is 5-7 cm (1).
Stickiness test: place 0.25 gram of lip cream
between the slides, and then add a load
weighing 1 kg on top of the slide for 5
minutes. After that, the weight was taken,
then the two slides were pulled using an 80-
gram load, and the time was recorded until
the slide was released. It is said to have
good adhesion when the object glass is
removed within 2-300 seconds (14).
Antioxidant Activity
Antioxidant activity of DrF-POMG was
determined by uv-vis spectrophotometer
UV-Vis (Shimadzu UV Mini-1240) using
DPPH method and vitamin C as a standard.
Determination of maximum wavelength
(λmaks) DPPH
The initial preparation is making a DPPH
solution, which is weighed at 10 mg and
then dissolved with 95% ethanol in 100 ml.
Pipette 4 ml of the solution, then add 2 ml
of 95% ethanol, and then determine the
maximum wavelength at 400–700 nm (15).
Determination of operating time
The solution was prepared by adding 2 mL
of DPPH and adding 4 mL of 95% ethanol,
then shaking until homogeneous. Then the
absorption is measured at the maximum
wavelength at 0, 10, 20, 30, 40, 50 and 60
minutes (15).
Preparation of standard vitamin C
solution
Vitamin C mains solution is prepared by
weighing 10 mg of vitamin C and then
dissolving it in 95% ethanol to 100 ml.
Dilute the solution so that 3 concentrations
are obtained, namely 2, 4, and 6 ppm.
Preparation of sample
The LCE5, LCE10 and LCE15 samples
were weighed as much as 10 mg and
dissolved in 95% ethanol up to 100 ml.
Each solution was diluted to obtain
concentrations of 40, 50 and 60 ppm.
Antioksidan measurement
Each 4 mL of vitamin C solution 2, 4, 6
ppm, added 2 mL of DPPH solution, shaken
until homogeneous, incubated 10 minutes
in a dark place. Measure the absorbance
using a UV-Vis spectrophotometer at λmax.
Each sample of LCE 5, LCE10 and LCE15
which had been diluted at a concentration
of 40, 50, 60 ppm was pipetted as much as
4 mL then added 2 mL of DPPH solution,
shaken until homogeneous, and incubated
for 10 minutes in a dark place. Measure the
absorbance using a UV-Vis
spectrophotometer at λmaks.
Data Analysis
This study uses data analysis
techniques with the Statview program
version 5.0 with Scheffe's testing. The data
analyzed were the results of the evaluation
of the preparations in the form of
measurements of pH, spreadability,
adhesion and antioxidant activity which
were then calculated on the average ± SD,
and the differences were compared
statistically. The results for the evaluation
of the preparation were compared with LC,
while the antioxidant activity test was
compared with vitamin C.
3. Result
Extraction of Red Dragon Fruit
Red dragon fruit was extracted using
the MAE (Microwave Assisted Extraction)
method with 95% ethanol and distilled
water (4:1) as the solvent to increase the
polarity of the sovent. Before MAE, the
simplisia was macerated by the solvent for
a night to wetting the simplisia. After
extraction was finished, the extract was
filtered with filter paper to obtain the
filtrate, then evaporation was carried out
with a rotary evaporator at 100 rpm to
evaporate the extraction solvent with a
temperature of 50ºC, to avoid damage to the
heat-resistant compounds.
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
351
Furthermore, the extract was
concentrated using a water bath to obtain a
thick extract. The extract obtained is brown
in color, this is because dragon fruit
contains anthocyanins which are a group of
flavonoids that have color stability that is
affected by light, temperature and also pH
(16). This method give the yield extract
about 44.12%. This results meets the
requirement because the yield extract
should be more than 10% (17).
A phytochemical screening were
examined to determine the secondary
metabolite contained in extracts. These
compounds identified by reagents that
provide the characteristics of each
secondary metabolite (18). The tests
included alkaloids, flavonoids, saponins,
steroids, triterpenoids and tannins. The
results of identifying secondary metabolites
of DrF extract can be seen in Table 2. DrF
extract contains alkaloid, flavonoid,
triterpenoids, saponin, and antochyanins,
but didn’t contained tannins and steroid
(18).
Table 2: Phytochemical screening of DrF Extract
Secondary metabolites
Results
Alkaloids
+
Flavonoids
+
Tannins
-
Triterpenoids
+
Steroids
-
Saponin
+
Antochyanins
+
Note: (+) = indicates present, (-) = indicates absent/not detected
Lip cream preparation for LC is white, LCE5 is light brown, LCE10 is brown and LCE15 is
dark brown.
Figure 1. Organoleptis appearance of LCE5, LCE10 dan LCE15
Organoleptic observation to see
whether the preparation were good or not,
that is seen from the shape, color and also
the smell is evenly distributed (12).
Organoleptically, the LC, LCE5, LCE10
and LCE15 preparations are semi-solid
(creamy) with a distinctive smell of roses.
LC which did not contain extract was white,
LCE5 was light brown, LCE10 and LCE15
were dark brown.
The pH measurement is carried out
to determine the acidity level of the
preparation so that it does not irritate when
applied to the lips. pH LC was 5.64±0.10,
LCE5 was 5.06±0.01, LCE10 was
5.05±0.03 and LCE15 was 5.04 ± 0.04.
This shows that the higher the
concentration of the extract will cause a
decrease in pH so that it becomes more
acidic. pH of LC preparations, LCE5.
LCE10 and LCE15 are in accordance with
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
352
the requirements for lip pH, namely 4.5-
6.5(13). The results of this pH measurement
show that all lip cream formulas are safe to
use. Homogeneity test was carried out to
determine whether there were coarse grains
in the preparation or not. Based on the tests
that have been carried out, all lip cream
preparations show that there are no coarse
grains in all formulas when placed between
glass objects. This result indicates that the
preparation is homogeneous and safe to
use. The spreadability test was carried
out to see how fast the lip cream can spread
on the lips when used so that it is easier to
apply. The scatter power results obtained
for LC = 5.14 ± 0.02 cm, LCE5 = 5.25 ±
0.02 cm, LCE10 = 5.33 ± 0.03 cm and
LCE15 = 5.34 ± 0.17 cm, so that the higher
the concentration of the extract used, the
more liquid the consistency and the easier it
is to apply. This spreadability test states that
all lipcream results meet the requirements,
namely 5-7 cm (12).
Antioxidant Activity
The DPPH method is used because
the method is simple, easy, only requires a
few samples, fast and sensitive. The
principle of this test is to measure the
damping power of an extract or sample
against DPPH free radicals. For
comparison, use vitamin C, because
vitamin C is a natural antioxidant that has
the function of capturing free radicals. At
the beginning of the test, a search for λmax
was carried out using a blank solution,
namely DPPH solution in 95% ethanol
(2:4) and scanning at λ = 400-700 nm using
a UV-Vis spectrophotometer (19).
The highest absorbance was
obtained at λmax = 519 nm of 0.988. This
λmax still meets the requirements for the
maximum DPPH wavelength of 515-530
nm (19). Operating time determination
starts from 0 minutes to 60 minutes. 0 to 10
minutes is the most stable time because it
produces the same absorbance value of
0.973 and will be used for the sample and
vitamin C incubation time, which is 10
minutes.
After determining the operating
time, proceed with measuring the
absorbance of vitamin C at concentrations
of 2, 4 and 6 ppm. For lip cream
preparations consisting of LCE5, LCE10
and LCE15 each was made with 3
concentrations namely 40, 50 and 60 ppm.
Sample solution of vitamin C and lip cream
each concentration added with DPPH
solution and also 95% ethanol which was
then incubated for 10 minutes beforehand.
If the sample solution contains a compound
that has a function as an antioxidant, when
it is reacted with DPPH, the initially dark
purple color will fade to bright purple or
yellow (5). After carrying out the
incubation process, the sample is inserted
into the UV-Vis spectrophotometer at a
maximum wavelength of 519 nm to
determine the absorbance value.
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
353
Table 3. Antioxidant activity
% inhibition was calculated by
using the equation Y = bx + a, with x is
concentration of each sample and Y is %
inhibition so that the IC50 value were
described in table 2. The IC50 for vitamin C,
LCE 5, LCE 10 and LCE 15 were 3.51± 0;
195.26 ± 0.40; 174.77 ± 0.34; and 168.27 ±
060, respectively. The higher the
concentration of the extract in LCE15 with
an extract concentration of 15%, resulting
the smaller IC50, which means the
antioxidant acvitity was more active to fight
the free radical of DPPH. Vitamin C has a
very strong antioxidant activity because the
IC50 is less than 50 ppm, while the
preparation has an IC50 value of more than
150 ppm which means it has moderate
antioxidant strength. In evaluation of the
antioxidant activity, temperature, light and
pH of the sample greatly affected the
results, and this can also be caused because
when the sample was made, the antioxidant
activity test is not immediately carried out
so that it affects the results of the
antioxidant activity. Based on the research
results obtained, it shows that red dragon
fruit extract can be used as a natural dye in
lip cream preparations by producing colors
from light brown to dark brown and has
antioxidant activity that can be used to
reduce free radicals.
4. Discussion
Red dragon fruit is extracted using
the MAE (Microwave Assisted Extraction)
method, because it only requires a short
extraction time, uses a small amount of
solvent and is easy. 95% ethanol is used as
a solvent because red dragon fruit contains
flavonoids which have a polarity close to
that of 95% ethanol and distilled water is
added to increase the polarity (20).
Organoleptic observation to see
whether the preparation that has been made
can be stated as good or not, that is seen
from the shape, color and also the smell is
evenly distributed (12). The results for LC,
LCE5, LCE10 and LCE15 preparations
were obtained in semi-solid (creamy) form
with a distinctive smell of roses. LC which
did not contain extract was white, LCE5
was light brown, LCE10 and LCE15 were
dark brown. so it can be concluded that this
lip cream preparation is good or meets the
requirements.
a. pH measurements were carried out to
determine the acidity level of the
preparation so as not to irritate it when
applied to the lips. The result of the pH
of LC, LCE5, LCE10 and LCE15
preparations were 5.64±0.10; 5.06±0.01;
5.05±0.03; and 5.04±0.04, respectively.
This result indicates that the higher the
I.Suharyani et al / Indo J Pharm 5 (2023) 347-356
354
concentration of the extract will produce
a more acidic pH value and this can also
be caused because this red dragon fruit
extract has an acidic pH of 5. The results
of this pH measurement are in
accordance with the requirements for lip
pH, namely 4.5-6.5 (13). The results of
this pH measurement show that all lip
cream formulas are safe to use.
b. Homogeneity test is carried out to find
out whether there are coarse grains in the
preparation or not. Based on the tests
that have been carried out, all lip cream
preparations show that there are no
coarse grains in all formulas when
placed between glass objects. This result
indicates that the preparation is
homogeneous and safe to use.
c. c. The spreadability test was carried out
to see how fast the lip cream can spread
on the lips when used so that it is easier
to apply. The spreadability results
obtained for LC, LCE5, LCE10 and
LCE15 preparations were 5.14±0.02 cm;
5.25±0.02 cm; 5.33±0.03 cm; and 5.34±
0.17 cm, respectively. The higher the
concentration of the extract used, the
more liquid the consistency and the
easier it is to apply. This spreadability
test stated that all the results of the
formula met the requirements, namely 5-
7 cm (12).
The DPPH method is used because
the method is simple, easy, requires only a
few samples, is fast and sensitive. The
principle of this test is to measure the
damping power of an extract or sample
against DPPH free radicals. For
comparison, use vitamin C, because
vitamin C is a natural antioxidant that has
the function of capturing free radicals.
The higher concentration of the
extract used as in LCE15 with an extract
concentration of 15%, the smaller the IC50
value obtained, which means the more
active the power of antioxidant compounds
that can be used to fight DPPH as free
radicals. Vitamin C has very strong
antioxidant activity because the IC50 is less
than 50 ppm, while the three formulations
have an IC50 value of more than 150 ppm
which means they have moderate
antioxidant strength. In testing the
antioxidant activity, temperature, light and
pH of the sample greatly affect the results
of the antioxidant activity test.
5. Conclusion
Based on the results of the research that has
been described, it can be concluded that red
dragon fruit extract can be used as a natural
coloring agent in lip cream preparations by
producing colors from light brown to dark
brown. The antioxidant activities for LCE5,
LCE10, and LCE15 were expressed in IC50
values, respectively, 195.26 ± 0.40
(moderate), 174.77 ± 0.34 (moderate),
168.27 ± 0.60 ( currently). LCE15 is the
best formula because it has a smaller IC50
value compared to LCE5 and LCE10,
which means that the lower the IC50 value,
the higher the antioxidant activity. Based
on the research results obtained, it shows
that red dragon fruit extract can be used as
a natural dye in lip cream preparations by
producing colors from light brown to dark
brown and has antioxidant activity that can
be used to reduce free radicals..
6. Acknowledgements
We would like to thank Sekolah Tinggi
Farmasi Muhammadiyah Cirebon for
supporting this study.
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